Immunological Relationship Research Articles

Infrared Fluorescence Imaging of Lymphatic Regeneration in Nonhuman Primate Facial Vascularized Composite Allografts

Clinical vascularized composite allografts (VCA), although performed with good success, have been characterized by rejection episodes and postoperative graft edema. We investigated lymphatic donor-recipient reconstitution and lymphatic regeneration in a nonhuman primate facial VCA model. Heterotopic partial face (n = 9) VCAs were performed in cynomolgus macaques. Grafts were monitored for rejection episodes and response to immunosuppressive therapies as previously described. Donor and recipient lymphatic channels were evaluated using a near-infrared handheld dual-channel light-emitting diode camera system capable of detecting fluorescence from indocyanine green injections. Graft lymphatic channels were serially evaluated from postoperative day 0 to 364. Preoperative imaging demonstrated superficial lymphatic anatomy similar to human anatomy. Initial resolution of facial allograft swelling coincided with superficial donor-recipient lymphatic channel reconstitution. Reconstitution occurred despite early acute rejection episodes in 2 animals. However, lymphatic channels demonstrated persistent functional and anatomic pathology, and graft edema never fully resolved. No differences in lymphatic channels were noted between grafts that developed transplant vasculopathy (n = 3) and those that did not (n = 6). Dynamic changes in patterns of lymphatic drainage were noted in 4 animals following withdrawal of immunosuppression. Donor-recipient lymphatic channel regeneration following VCA did not result in resolution of edema. Technical causes of graft edema may be overcome with alternative surgical techniques, allowing for direct investigation of the immunologic relationship between VCA graft edema and rejection responses. Mechanisms and timing of dynamic donor-recipient lymphatic channel relationships can be evaluated using fluorescent imaging systems to better define the immunologic role of lymphatic channels in VCA engraftment and rejection responses, which may have direct clinical implications.

Histologic and Immunohistochemical Analyses of α-Fetoprotein—Producing Cancer of the Stomach

As the histogenesis and development of α-fetoprotein-producing gastric cancer (AFPGC) have not yet been elucidated, we analyzed the histologic and immunologic relationship between the histologic type of the mucosal lesion considered to be the primary lesion, and that of its invasive lesion, in 36 cases of AFPGC. We reviewed 23 AFPGCs with mucosal lesions (1 mucosal and 22 submucosal or deeper invasive tumors) among 36 AFPGCs that had been resected endoscopically or surgically between 1970 and 2005. AFPGC was defined as a tumor showing immunohistochemical positivity for either α-fetoprotein (AFP) or glypican-3. Histologic types were divided into hepatoid (HPT), enteroblastic (ENT), yolk sac tumor, and common (COM) adenocarcinoma type. The tumor phenotypes were classified into gastric, gastrointestinal, and intestinal types on the basis of immunohistochemical analysis. Among the histologic types of mucosal lesions, the COM and ENT mixed type was observed in 65.2% of cases (15/23 patients), COM alone in 26.1% (6/23), and ENT alone in 8.7% (2/23) of cases. Among the invasive lesions, 16 cases (72.7%) were HPT. Both AFP and glypican-3 were positive in 60.9% (14/23) of mucosal lesions and in 90.9% (20/22) of invasive lesions. With regard to phenotypic expression, 82.6% (19/23) of mucosal lesions were the intestinal type, compared with 95.5% (21/22) of invasive lesions. These findings suggest that many cases of AFPGC develop as COM or ENT in the mucosa, which differentiate into ENT and HPT during the process of tumor invasion and proliferation, acquiring AFP production ability.

진균과 식물의 Phenylalanine Ammonia-lyase 그리고 세균의 Histidine Ammonia-lyase 간의 면역학적 관계 분석

진균의 phenylalanine ammonia-lyase(PAL)와 식물의 PAL간 그리고 PAL과 histidine ammonia-lyase (HAL)간의 구조적 특성 관계에 대한 이해를 얻기 위해 옥수수병원균인 깜부기균(Ustilago maydis)의 PAL에 대한 면역학적 분석을 수행하였다. 진균의 PAL 항체와 식물의 PAL 항체를 이용하여 분석하였을 때 실험한 모든 종의 PAL 간에 교차반응이 나타났다. 알팔파 PAL 항체와 포플라 PAL 항체 모두 식물 PAL은 강하게 인식하였으나 진균의 PAL은 약하게 인식하였다. U. maydis PAL은 Rhodotorula glutinis 효모의 PAL만 약하게 인식하였다. U. maydis PAL은 식물의 PAL에 대해서 낮은 친화성을 보였으나 Pseudomonas 세균의 HAL에 대해서는 강한친화성을 보였다. 2종의 식물 PAL 항체들 또한 Pseudomonas 세균의 HAL에 대해 강한 친화성을 보였다. 진균과 식물의 PAL 항체는 PAL 효소의 활성 저해를 나타냈으며 세균의 HAL 효소의 활성에 대해서도 중도적 저해를 나타냈다. 세균의 HAL 항체는 식물과 효모와 Ustilago PAL 중에 Ustilago PAL 활성만 저해하였다. 식물의 PAL과 진균의 PAL 모두 효모의 PAL 활성은 저해하지 못했다. 본 연구는 PAL과 HAL간에 면역학적 관계가 있음을 처음으로 보고한다. Phenylalanine ammonia-lyase (PAL) from the maize pathogen Ustilago maydis was analysed immunologically to obtain insights into the structural relationships between plant PAL and fungal PAL and between PAL and histidine ammonia-lyase (HAL). Cross-reactivity was found among all the PAL proteins from different species tested, using antibodies raised against both plant and fungal PALs. Both anti-Alfalfa and anti-popular PAL antibodies strongly recognized plant PALs but only weakly recognized fungal PALs. Antibodies raised against U. maydis PAL only weakly recognized the Rhodotorula glutinis yeast PAL. The anti-U. maydis PAL antibodies showed low affinity for the plant PALs but they bound strongly to Pseudomonas bacterial HAL. Significant cross-reactivity between the two plant PAL antibodies and the bacterial HAL was also observed. Both the anti-Ustilago PAL and the anti-poplar PAL antibodies displayed similar enzyme inhibition patterns, including moderate inhibition of bacterial HAL activity. However, the bacterial HAL antibody inhibited only Ustilago PAL. The PAL and HAL antibodies tested showed no inhibition against yeast PAL. This is first report on the immunological relationships between PAL and HAL.

Immunologic Regulation in Pregnancy: From Mechanism to Therapeutic Strategy for Immunomodulation

The immunologic interaction between the fetus and the mother is a paradoxical communication that is regulated by fetal antigen presentation and/or by recognition of and reaction to these antigens by the maternal immune system. There have been significant advances in understanding of abnormalities in the maternal-fetal immunologic relationship in the placental bed that can lead to pregnancy disorders. Moreover, immunologic recognition of pregnancy is vital for the maintenance of gestation, and inadequate recognition of fetal antigens may cause abortion. In this paper, we illustrate the complex immunologic aspects of human reproduction in terms of the role of human leukocyte antigen (HLA), immune cells, cytokines and chemokines, and the balance of immunity in pregnancy. In addition, we review the immunologic processes of human reproduction and the current immunologic therapeutic strategies for pathological disorders of pregnancy.

Molecular cloning, heterologous expression and functional characterization of a novel translationally-controlled tumor protein (TCTP) family member from Loxosceles intermedia (brown spider) venom

Envenoming with brown spiders (Loxosceles genus) is common throughout the world. Cutaneous symptoms following spider bite accidents include dermonecrosis, erythema, itching and pain. In some cases, accidents can cause hypersensibility or even allergic reactions. These responses could be associated with histaminergic events, such as an increase in vascular permeability and vasodilatation. A protein that may be related to the effects of spider venom was identified from a previously obtained cDNA library of the L. intermedia venom gland. The amino acid sequence of this protein is homologous to proteins from the TCTP (translationally-controlled tumor protein) family, which are extracellular histamine-releasing factors (HRF) that are associated with the allergic reactions to parasites. Herein, we described the cloning, heterologous expression, purification and functional characterization of a novel member of the TCTP family from the Loxosceles intermedia venom gland. This recombinant protein, named LiRecTCTP, causes edema, enhances vascular permeability and is likely related to the inflammatory activity of the venom. Moreover, LiRecTCTP presents an immunological relationship with mammalian TCTPs.

Antiphospholipid syndrome and rheumatic fever: a case spanning three decades of changing concepts and common immunological mechanisms

We present a case of primary antiphospholipid syndrome (APS), initially diagnosed as acute rheumatic fever, resulting in severe mitral valve incompetence. This case raises questions of the specificity of the Jones diagnostic criteria for rheumatic fever in a population where it is infrequently encountered. There are similarities in clinical, pathological and echocardiographic presentations between rheumatic fever and APS, in addition to common immunological mechanisms. Our case highlights the possibility that rather than rheumatic fever being primarily responsible for her recurrent attacks of chorea and arthritis, the streptococcal infections in our patient occurred either in the setting of underlying antiphospholipid antibodies ('second hit' phenomenon), or may have triggered the development of pathogenic antibodies (molecular mimicry), subsequently leading to the clinical evolution of APS. During the three decades of our patient and her recurrent problems, there has been an evolving knowledge of the mechanisms of APS and rheumatic fever, allowing us to extend our understanding beyond symptoms and syndromes, to a better realization of the underlying immunological relationship between the two.

Recent advances in understanding immunology of reproductive failure

Aspects of the immunological relationship between mother and conceptus still remain a mystery, although the recent advances in molecular biology have enlightened some of the parameters that participate in fetomaternal cross-talk during implantation. The atypical expression of major histocompatibility complex (MHC), the specific role of some hormones and cytokines, as well as the temporal and spatial distributions of uterine natural killer cells, represent substantive parameters of fetomaternal immunotolerance during implantation. Although human maternal and fetal immunology is difficult to investigate, aberrant immune responses and an imbalanced cytokine network may be related to infertility, implantation failures after IVF and recurrent pregnancy losses. In this review, immunological and interacting factors involved in human reproductive failure are summarized and critically evaluated.

Functional immunoglobulin E cross‐reactivity between Pas n 1 of Bahia grass pollen and other group 1 grass pollen allergens

Grass pollens are major triggers of allergic rhinitis and asthma, but the immunological relationships between pollen allergens of the subtropical Bahia grass, Paspalum notatum, and temperate grasses are unresolved. To assess serum IgE cross-reactivity between subtropical P. notatum and temperate Lolium perenne (Ryegrass) pollen allergens. Serum IgE reactivities of grass pollen-allergic patients with P. notatum, L. perenne and Cynodon dactylon (Bermuda grass) pollen extracts and their respective purified group 1 allergens, Pas n 1, Lol p 1 and Cyn d 1, were compared by immunoblotting, ELISA and basophil activation. In a cohort of 51 patients from a temperate region, a high frequency of IgE reactivity with each grass pollen was detected, but reactivity with L. perenne pollen was substantially greater than with P. notatum and C. dactylon pollen. Similarly, serum IgE reactivity with Lol p 1 was greater than with Pas n 1 or Cyn d 1. For seven of eight sera studied in detail, asymmetric serum IgE cross-reactivity was observed; L. perenne pollen inhibited IgE reactivity with P. notatum pollen but not the converse, and IgE reactivity with Pas n 1 was inhibited by Lol p 1 but IgE reactivity with Lol p 1 was not inhibited by Pas n 1 or Cyn d 1. Importantly, P. notatum pollen and Pas n 1 activated basophils in grass pollen-allergic patients from a temperate region, although stimulation was greater by pollen of L. perenne than P. notatum or C. dactylon, and by Lol p 1 than Pas n 1 or Cyn d 1. In contrast, a cohort of 47 patients from a subtropical region showed similar IgE reactivity with P. notatum and L. perenne pollen, and reciprocal cross-inhibition of IgE reactivity between L. perenne and P. notatum. Pollen allergens of the subtropical P. notatum, including Pas n 1, show clinically relevant IgE cross-reactivity with pollen allergens of L. perenne but also species-specific IgE reactivity.

Structural and immunologic cross-reactivity among filarial and mite tropomyosin: Implications for the hygiene hypothesis

The hygiene hypothesis suggests that parasitic infection modulates host immune responses and decreases atopy. Other data suggest parasitic infections may induce allergic responsiveness. To assess the structural and immunologic relationships between the known Dermatophagoides pteronyssinus (Der p 10) tropomyosin allergen and filarial tropomyosin of Onchocerca volvulus (OvTrop). The molecular, structural, and immunologic relationships between OvTrop and Der p 10 were compared. Levels of OvTrop-specific and Der p 10-specific IgE, IgG, and IgG₄ in sera of filaria-infected and filarial-uninfected D pteronyssinus-atopic individuals were compared, as were the responses in nonhuman primates infected with the filarial parasite Loa loa. Cross-reactivity was compared by antigen-mediated depletion assays and functionality by passive basophil sensitization. Filarial and mite tropomyosins were very similar, with 72% identity at the amino acid level, and overlapping predicted 3-dimensional structures. The prevalence of IgE and IgG to Der p 10 was increased in filaria-infected individuals compared with uninfected subjects. There was a strong correlation between serum levels of Ov- and Der p 10-tropomyosin-specific IgE, IgG, and IgG₄ (P < .0001; r > 0.79). Preincubation of sera from anti-Der p 10-positive subjects with OvTrop completely depleted IgE, IgG, and IgG₄ anti-Der p 10. Basophils sensitized with sera from individuals allergic to Der p 10 released histamine similarly when triggered with OvTrop or Der p 10. Primates experimentally infected with L loa developed IgE that cross-reacted with Der p 10. Filarial infection induces strong cross-reactive antitropomyosin antibody responses that may affect sensitization and regulation of allergic reactivity.

Toxocara infection and diminished lung function in a nationally representative sample from the United States population

The relevance of parasitic infection for the increasing incidence of asthma is a topic of considerable debate. Large population-based studies examining the association between helminth infection and specific measures of lung function in humans are lacking. This report sought to examine this association by exploring the differences in forced expiratory volume in 1s (FEV1) among participants with and without infection with Toxocara spp. using data from the Third National Health and Nutrition Examination Survey, undertaken by the United States Department of Health and Human Services, during 1988–1994. The results showed a significant association between diminished lung function and previous infection with Toxocara spp. Those with antibody evidence of Toxocara infection displayed FEV1 that was 105.3mL less than those without previous infection. This relationship persisted while controlling for age, sex, education level, BMI, smoking status, ethnicity, immigration, rural residence and dog ownership (fully-adjusted difference=73mL). These findings suggest diminished lung function in the presence of Toxocara infection and illustrate the urgent need for longitudinal data to more clearly define the immunological relationship with helminth infection and its potential influence on lung function.

Helminth secretions induce de novo T cell Foxp3 expression and regulatory function through the TGF-β pathway

Foxp3-expressing regulatory T (T reg) cells have been implicated in parasite-driven inhibition of host immunity during chronic infection. We addressed whether parasites can directly induce T reg cells. Foxp3 expression was stimulated in naive Foxp3⁻ T cells in mice infected with the intestinal helminth Heligmosomoides polygyrus. In vitro, parasite-secreted proteins (termed H. polygyrus excretory-secretory antigen [HES]) induced de novo Foxp3 expression in fluorescence-sorted Foxp3⁻ splenocytes from Foxp3-green fluorescent protein reporter mice. HES-induced T reg cells suppressed both in vitro effector cell proliferation and in vivo allergic airway inflammation. HES ligated the transforming growth factor (TGF) β receptor and promoted Smad2/3 phosphorylation. Foxp3 induction by HES was lost in dominant-negative TGF-βRII cells and was abolished by the TGF-β signaling inhibitor SB431542. This inhibitor also reduced worm burdens in H. polygyrus-infected mice. HES induced IL-17 in the presence of IL-6 but did not promote Th1 or Th2 development under any conditions. Importantly, antibody to mammalian TGF-β did not recognize HES, whereas antisera that inhibited HES did not affect TGF-β. Foxp3 was also induced by secreted products of Teladorsagia circumcincta, a related nematode which is widespread in ruminant animals. We have therefore identified a novel pathway through which helminth parasites may stimulate T reg cells, which is likely to be a key part of the parasite's immunological relationship with the host.

Rapid identification of vector-borne flaviviruses by mass spectrometry

Flaviviruses are a highly diverse group of RNA viruses classified within the genus Flavivirus, family Flaviviridae. Most flaviviruses are arthropod-borne, requiring a mosquito or tick vector. Several flaviviruses are highly pathogenic to humans; however, their high genetic diversity and immunological relatedness makes them extremely challenging to diagnose. In this study, we developed and evaluated a broad-range Flavivirus assay designed to detect both tick- and mosquito-borne flaviviruses by using RT-PCR/electrospray ionization mass spectrometry (RT-PCR/ESI-MS) on the Ibis T5000 platform. The assay was evaluated with a panel of 13 different flaviviruses. All samples were correctly identified to the species level. To determine the limit of detection for the mosquito-borne primer sets, serial dilutions of RNA from West Nile virus (WNV) were assayed and could be detected down to an equivalent viral titer of 0.2 plaque-forming units/mL. Analysis of flaviviruses in their natural biological background included testing Aedes aegypti mosquitoes that were laboratory-infected with dengue-1 virus. The assay accurately identified the virus within infected mosquitoes, and we determined the average viral genome per mosquito to be 2.0 × 10 6. Using human blood, serum, and urine spiked with WNV and mouse blood and brain tissues from Karshi virus-infected mice, we showed that these clinical matrices did not inhibit the detection of these viruses. Finally, we used the assay to test field-collected Ixodes scapularis ticks collected from sites in New York and Connecticut. We found 16/322 (5% infection rate) ticks positive for deer tick virus, a subtype of Powassan virus. In summary, we developed a single high-throughput Flavivirus assay that could detect multiple tick- and mosquito-borne flaviviruses and thus provides a new analytical tool for their medical diagnosis and epidemiological surveillance.

Immune responses of the domestic fowl to Dermanyssus gallinae under laboratory conditions

There appear to be few reports in the literature regarding the host-poultry red mite (Dermanyssus gallinae) immunological relationship, despite the negative impact D. gallinae can have on both bird welfare and egg production, as well as its potential to act as a reservoir of zoonotic infections. The current study investigated the effect of either continuous infestation (CI) for 22 days or repeated infestation (RI) for four 24-h periods 7 days apart with D. gallinae on humoral immunity (IgM and IgY) and Th1/Th2 cytokine mRNA expression in peripheral blood mononuclear cells (PBMC) compared to non-infested controls. Serum IgY levels and IgM concentration were significantly higher in CI than RI and control birds although Th1 and Th2 mRNA expression in PBMC did not differ significantly between groups. D. gallinae appeared to modify reproductive behaviour and progeny survival following successive feeding events. In the RI group, the proportion of eggs/mite was significantly higher (P < 0.05) after first infestation than later infestations while larval/nymphal mortality was significantly higher (P < 0.05) after the first two infestations than later infestations. These data suggest that D. gallinae might adopt a feeding strategy of minimal host interference while D. gallinae could determine host immune status via nymphal/larval survival rates. Further research is required to better understand the host immunomodulation or avoidance strategy of D. gallinae as well as whether the mite is able to determine host immunocompetence perhaps using progeny survival.

Paraneoplastic Neurologic Syndromes - An Update on Current Understanding and Future Perspectives

Paraneoplastic neurological syndromes (PNS) are remote effects of tumours on the nervous system. They can strike at single or at multiple sites of the central nervous system (CNS) and the peripheral nervous system, and often appear before the detection of cancer. PNS can be disabling and debilitating and may be either an additional burden to cancer or the cause of death. The PNS Euronetwork group has collected a series of approximately 1,000 patients in several European centres. This study is the largest systematic series of patients with PNS and, for the first time, can answer questions about the most frequent PNS, their detailed symptoms, associated antibodies and the types of underlying tumours. The clinical course and laboratory findings for many PNS suggest an autoimmune pathogenesis; however research into this heterogeneous immunological relationship has been evolving over recent decades. The classical syndromes are antigen-target-oriented syndromes such as myasthenia gravis (MG), Lambert-Eaton myasthenic syndrome (LEMS) and ion channelmediated diseases. Onconeuronal antibodies constitute a large group of PNS, characterised by the appearance of specific antibodies, defined clinical signs and often an association with specific tumours. In recent years, a new group of antibodies directed at surface antigens as receptors has been identified. Finally there is a long list of ‘other’ PNS, which are evident to clinicians but which have no pathogenetic explanation. Examples include the mild terminal neuropathies and sarcopoenia in cancer patients. In addition to the emerging classification based on pathophysiology, other new syndromes and symptoms have appeared, including apnoea in brainstem encephalitis, a neuropsychiatric spectrum of limbic encephalitis and increased knowledge about LEMS. Two important aspects warrant attention: some PNS respond to therapy and not all paraneoplastic-like syndromes are tumour related. This view is based on the current understanding of immune pathogenesis and on the enlarged spectrum of PNS.

Split immunological tolerance to trophoblast

Split immunological tolerance refers to states in which an individual is capable of mounting certain types of immune responses to a particular antigenic challenge, but is tolerant of the same antigen in other compartments of the immune system. This concept is applicable to the immunological relationship between mother and fetus, and particularly relevant in equine pregnancy. In pregnant mares, antibody responses to paternal foreign Major Histocompatibility Complex class I antigens are robust, while anti-paternal cytotoxic T cell responses are diminished compared to those mounted by non-pregnant mares. Here, we compared the distribution of the major lymphocyte subsets, the percentage of lymphocytes expressing Interferon Gamma (IFNG) and Interleukin 4 (IL4) and the level of expression of the immunoregulatory transcription factor FOXP3 between pregnant and non-pregnant mares, and between peripheral blood and the endometrium during pregnancy. In a cohort of mares in which peripheral blood lymphocytes were tested during early pregnancy and in the non-pregnant state, there were only slight changes observed during pregnancy. In contrast, comparison of peripheral blood lymphocytes with lymphocytes isolated from the endometrial cups of pregnant mares revealed striking differences in lymphocyte sub-populations. The endometrial cups contained higher numbers of IFNG+ lymphocytes, and lower numbers of lymphocytes expressing IL4. The endometrial cup lymphocytes also had higher numbers of FOXP3+ cells compared to peripheral blood lymphocytes. Taken together, these results strengthen the evidence for a state of split tolerance to trophoblast, and furthermore define sharp differences in immune reactivity during equine pregnancy between peripheral blood lymphocytes and lymphocytes at the maternal-fetal interface.

Paraneoplastic Neurologic Syndromes—An Update on Current Understanding and Future Perspectives

Paraneoplastic neurological syndromes (PNS) are remote effects of tumors on the nervous system. They can strike at single or at multiple sites of the central nervous system (CNS) and the peripheral nervous system, and often appear before the detection of cancer. PNS can be disabling and debilitating and may be either an additional burden to cancer or the cause of death. The PNS Euronetwork group has collected a series of approximately 1,000 patients in several European centers. This study is the largest systematic series of patients with PNS and, for the first time, can answer questions about the most frequent PNS, their detailed symptoms, associated antibodies, and the types of underlying tumors. The clinical course and laboratory findings for many PNS suggest an autoimmune pathogenesis; however, research into this heterogeneous immunological relationship has been evolving over recent decades. The classic syndromes are antigen-target-oriented syndromes such as myasthenia gravis (MG), Lambert-Eaton myasthenic syndrome (LEMS), and ion channel-mediated diseases. Onconeuronal antibodies constitute a large group of PNS, characterized by the appearance of specific antibodies, defined clinical signs and often an association with specific tumors. In recent years, a new group of antibodies directed at surface antigens as receptors has been identified. Finally, there is a long list of ‘other’ PNS, which are evident to clinicians but which have no pathogenetic explanation. Examples include the mild terminal neuropathies and sarcopenia in cancer patients. In addition to the emerging classification based on pathophysiology, other new syndromes and symptoms have appeared, including apnea in brainstem encephalitis, a neuropsychiatric spectrum of limbic encephalitis, and increased knowledge about LEMS. Two important aspects warrant attention: some PNS respond to therapy and not all paraneoplastic-like syndromes are tumor related. This view is based on the current understanding of immune pathogenesis and on the enlarged spectrum of PNS.

Determinants of the Relationship between Cytokine Production in Pregnant Women and Their Infants

Exposure to environmental factors during fetal life and infancy is thought to play an important role in the early development of innate and adaptive immunity. The immunological relationship between mother and infant and the effect that environmental exposures have during pregnancy and early childhood have not been studied extensively. Here the production of cytokines was measured in 146 pairs of mothers and their 2- month-old infants. The effect of place of residence, socio-economic variables, parasitic infections as well as maternal and child characteristics on measured cytokine production was determined. Mothers producing high levels of IL-10, IFN-γ and IL-5 were more likely to have infants who also produced high levels of these cytokines either spontaneously (OR 2.6(95%CI 1.2–5.4), OR 2.9(CI 1.3–6.6), OR 11.2(CI 4.6–27.2), respectively) or in response to PHA (IL-10: OR 3.0(CI 1.4–6.6), IFN-γ: OR 2.0(CI 1.0–4.2), respectively) even after adjustment for potential confounding variables. This was not the case for TNF-α. In response to LPS, place of residence was a strong determinant of infant IL-10 (OR 0.2(CI 0.1–0.9)) and TNF-α (OR 0.3(CI 0.1–0.9)) production. Maternal protozoan infections was independently associated with reduced infant IL10 in response to PHA and to LPS as well as reduced TNF-α and IFN-γ in response to PHA. These results indicate strong relationship between maternal and infant's cellular immune responses even after taking into account many environmental influences that could affect infant's response directly or indirectly through uterine microenvironment. However, place of residence and intestinal infections may still directly affect the immune responses of the infant. Taken together, the study provides evidence for imprinted cytokine responses of an infant which may have implications for their reaction to incoming antigens, warranting further investigation into the role that genetics or epigenetics play in shaping the cytokine response by an infant to self or external antigens.

Strepsirhine dichotomy from a human perspective (Primates: Lorisoidea, Lemuroidea)

Using monospecific, polyclonal antisera against 69 human plasma proteins, 128 antigenic determinants from 40 cross-reacting homologues were characterized in representatives of the prosimian genera Lemur, Eulemur, Varecia (Lemuridae) and Otolemur (Galagidae). Seventeen determinants from 16 different proteins were absent in homologues of the gagalo but were shared by lemurs and several platyrrhines, cercopithecids, and hominoids. Smaller locus samples for potto(Perodicticus potto) and slow loris (Nycticebus coucang) confirmed the more distant immunological relationship of lorises than of lemurs to anthropoids. If evolutionary rates are constant and equal in lorisiform and lemuriform prosimians, this patern of character distribution indicates strepsirhine paraphyly, lorises diverging earlier (possibly some 6 times 106 years) than lemurs from anthropoid ancestors. If this is so, lorises rather than true lemurs should be elected to root the polarity of character evolution in Primates. As an alternative, galagine proteins evolve more rapidly than lemurine homologues.

Involvement of glycans in the immunological cross-reaction between α-macroglobulin and hemocyanin of the gastropod Helix pomatia

By tandem-crossed immunoelectrophoresis and ELISA experiments an immunological relationship was observed between α-macroglobulin (αM) and hemocyanin (Hc) of the terrestrial snail Helix pomatia. Both glycoproteins occur in the hemolymph: αM (minor component) as a specific proteinase inhibitor, Hc (consisting of three components: α D- HpH, α N- HpH and β- HpH) as oxygen transport protein. The cross-reaction was found to be correlated with glycosylation. (i) With β- HpH, which is richer in carbohydrates than α D- HpH and α N- HpH, mainly due to a higher 3- O-methyl- d-galactose content, the cross-reaction with HpαM was highest. (ii) From the 8 functional units, designated a– h, isolated from β- HpH, two that lack carbohydrates ( c and f) were not recognized by antibodies against HpαM, while the six glycosylated ones were strongly cross-reacting. The nearly complete loss of the cross-reactivity upon deglycosylation of functional units d and g and the inhibition in competitive ELISA experiments by glycopeptides isolated from both β- HpH and HpαM are further evidence that glycans are involved in the immunological relationship between HpH and HpαM. Carbohydrate analyses indicated that the glycan structures present on HpαM are very similar (or identical) to those found on HpH, suggesting that glycans are common epitopes on both proteins. Especially d-xylose and 3- O-methyl- d-galactose seem to be responsible for the cross-reactivity since the α-macroglobulin and hemocyanin of the cephalopod Sepia officinalis, which lack these two monosaccharides in their glycan structures, do not immunologically cross-react.

Expression of endogenous retrovirus ev/J gp85 gene and analysis of its immunoreactivity in comparison with exogenous viral protein

The envelope gene gp85 of ev/J, a new family of endogenous avian retroviral sequences identified recently, has the most extensive nucleotide sequence identity ever described with ALV-J avian leukosis virus. This report described expression of ev/J envelope gene gp85 derived from commercial meat-type chicken using the Invitrogen Bac-to-Bac baculovirus expression system. The antigenicity and immunoreactivity of the recombinant endogenous gp85 gene product (SU) were analyzed by indirect immunofluorescence, Western blot, indirect and blocking Enzyme-Linked ImmunoSorbent Assay (ELISA) using JE9 monoclonal antibody (MAb) against the envelope protein of ALV-J (ADOL-4817), positive mouse antiserum against the ev/J gp85 SU and sera from chicken naturally infected with ALV-J. The results showed that the ev/J gp85 SU can bind specifically to JE9 MAb and antiserum from chicken naturally infected with ALV-J, and the binding reactivity between exogenous ALV-J gp85 SU and natural positive chicken serum against exogenous ALV-J can be blocked by positive mouse serum against the ev/J gp85 SU. It is concluded that recombinant endogenous gp85 gene product (SU) has close immunological relatedness to the envelope protein of exogenous ALV-J (ADOL-4817 and IMC10200 strain).